脲对果菠萝蛋白酶活力与构象的影响

本文用荧光光谱,紫外差示光谱和CD谱研究果菠萝蛋白酶在不同浓度的脲溶液中的构象及酶活力的变化情况。酶的荧光强度随脲浓度增大而明显增加,8mol/L脲使荧光强度增强65％,发射峰出现红移。差示谱表明在232nm和288nm出现二个正峰,它们均随脲浓度增大而加剧,前者与主链构象变化有关,而后者与生色基团(Trp、Tyr)的微环境变化相关。CD谱表明:天然酶在208nm和225nm处有二个负峰,脲变性后,225nm的负峰基本上不随脲浓度增大而变化,但208nm峰则明显发生变化并逐渐出现红移,6mol/L以上此峰则完全消失。     

维生素E琥珀酸酯的合成及表征

目的:合成维生素E琥珀酸酯并对其进行表征.方法:以d,1-α-生育酚和丁二酸酐为原料,吡啶为溶媒,合成了维生素E琥珀酸酯.采用紫外光谱法、红外光谱法、核磁共振氢谱和差示扫描量热分析对产物进行表征.结果:经验证,成功合成了维生素E琥珀酸酯.结论:成功合成了维生素E琥珀酸酯.     

蛋白质修饰剂对盐藻光合作用的影响

经蛋白质化学修饰剂N-溴代琥珀酰亚胺（NBS）、丁二酮（BTO）和对-氯汞苯甲酸（ρCMB）处理的盐藻细胞光合速率下降,0.17mmol/L的ρCMB和0.07mmol／L的NBS可完全抑制光合放氧。在藻细胞的可见光（400—700nm）区吸收光谱中,三种修饰剂都降低了整个波段的吸收。在两个主要吸收峰中,678nm吸收值的下降略大于436nm的下降。在紫外光谱区（200—300nm）,ρCMB和BTD使原吸收峰（203nm）值明显降低,NBS处理使吸收峰红移13nm。细胞胀破后紫外光谱出现更显著变化,峰位移至223nm（BTD）、250nm（NBS）,或至214—237nm而呈一个宽的平台（ρCMB）。紫外差示吸收光谱显示210nm的负峰;随修饰剂浓度增大,负差示峰可移到225nm（NBS）、245．5nm（ρCMB）和212nm（BTD）。     

牛红细胞超氧化物歧化酶活性中心的紫外光谱研究

应用差示分光光度法研究了牛红细胞Ｃｕ２Ｚｎ２ＳＯＤ的紫外光谱,归属和讨论了酶活性中心金属离子与配体间全部电荷转移谱带,给出了相应的配体轨道光学电负性,特别研究了涉及Ｚｎ２＋的电荷转移谱带．     

使用差示扫描量热仪测定抗冻蛋白热滞活性方法的研究

抗冻蛋白因具有独特的抗冻活性而被研究者广泛关注。但是,目前抗冻活性的检测没有一个标准的、统一的检测方法,这严重制约了该方面的研究进展。作者详细研究了采用差示扫描量热仪测定样品热滞活性的方法,并对该方法的稳定性、专一性和精密度进行评价。结果显示,采用差示扫描量热仪测定样品的热滞活性具有较高的稳定性、重复性和精密度。因此,差示扫描量热仪法可以作为一种通用的方法进行抗冻蛋白热滞活性的检测。     

差示扫描量热法在红细胞研究中的应用

差示扫描量热法(DSC)是在程序控制温度下,测量试样和参比物能量差随温度(时间)的变化关系的一种技术。它通过捕捉细胞升降温过程中的吸放热现象,利用热学参数(焓变、比热、相变温度)来表征其间细胞结构的变化情况。量热学方法与结构分析方法相结合,有助于深入剖析细胞的生理变化。本文从预测低温保存过程中最佳降温速度及分析细胞膜损伤机理两方面探讨差示扫描量热法在红细胞研究中的应用。     

稳恒磁场对两种不同构象状态的离体牛肝过氧化氢酶的生物学效应研究

通过活力测定,紫外差光谱,多维荧光光谱及差示扫描量热分析,研究了0.23～0.61T稳恒磁场对两种不同构象状态的离体牛肝过氧化氢酶的生物学效应,被选择用于研究的酶的构象状态分别为4℃的钝化状态和25℃的活化状态,二者具有明显不同的构象,4℃时,酚分子处于钝化状态,经0.23～0.61T稳恒磁场处理不同的时间后,几乎不表现出任何磁生物学效应;25℃时,酶分子处于活化状态,经磁场处理后,表现出明显的磁生物学效应;酶活力增加,同时构象发生变化。构象变化导致λ210～310nm紫外差光谱的出现,荧光偏振度的增加,在λ330nm荧光发射峰发射强度的改变及差示扫描量热曲线的产生,研究结果表明：不同的初始构象状态可能是产生不同磁效应的根本原因。     

渗透处理对麻栎和栓皮栎种子生物学特性的影响

本研究旨在探究渗透处理对麻栎(Quercus acutissima)和栓皮栎(Q.variabilis)种子生物学特性的影响,分析渗透处理过程中种子内部自由水含量的变化,为麻栎和栓皮栎种质资源开发利用提供理论参考。本研究分别使用浓度为0%、 25%、 50%和75%的PEG-6000溶液对麻栎和栓皮栎种子进行渗透处理,绘制种子含水率变化曲线图,并对萌发指标进行评测;基于差示扫描量热(differential scanning calorimetry, DSC)技术分析不同浓度PEG处理后种子热力学特征变化;使用扫描电子显微镜观察不同浓度PEG处理后胚轴细胞超微结构变化。结果显示去除种皮后的麻栎和栓皮栎种子平均含水率(40.56%、 44.31%)和发芽率(97.77%、 95.55%)均高于带种皮种子的含水率(34.94%、 33.51%)和发芽率(75.00%、 77.78%), 50%PEG对麻栎和栓皮栎种子萌发促进效果最好,75%PEG的效果最差;胚轴结晶起始温度、峰值温度及热焓值随PEG浓度的升高均呈规律性下降趋势;扫描电镜观测结果发现高浓度PEG导致细胞降解,部分细胞出现皱缩...     

菠萝叶片PEP羧激酶与底物结合时构象的变化

菠萝叶片PEP羧激酶与底物OAA和ATP及配基Mn~(2+)等结合时引起紫外差示吸收光谱峰位和方向上的变化。OAA与酶结合诱导产生的差示吸收光谱在268—280nm处有一个宽负峰。ATP与酶结合出现两个差示负峰(242.5和273.5nm)。双底物OAA和ATP同时与酶结合,光谱上呈现252nm和268nm两个峰。Mn~(2+)不论与ATP或与ATP+OAA一起与酶反应时,皆使原来的峰位漂移,且正负方向逆转。酶蛋白在323nm有最大的荧光发射。OAA引起荧光发射强度增大,ATP及ATP+Mn~(2+)则减弱荧光发射。Mn~(2+)与OAA及ATP的复合效应导致荧光强度进一步减弱。     

菠萝叶片PEP羧激酶与底物结合时构象的变化

菠萝叶片PEP羧激酶与底物OAA和ATP及配基Mn^2 等结合时引起紫外差示吸收光谱峰位和方向上的变化。OAA与酶结合诱导产生的差示吸收光谱在268—280mm处有一个宽负峰。ATP与酶结合出现两个差示负峰(242．5和273．5nm)。双底物OAA和ATP同时与酶结合，光谱上呈现252nm和268nm两个峰。Mn^2 不论与ATP或与ATP OAA一起与酶反应时，皆使原来的峰位漂移，且正负方向逆转。酶蛋白在323nm有最大的荧光发射。OAA引起荧光发射强度增大，ATP及ATP Mn^2 则减弱荧光发射。Mn^2 与OAA及ATP的复合效应导致荧光强度进一步减弱。     

艾氏腹水癌患鼠腹水中一种高分子量DNA结合蛋白的分离

 本文利用免疫吸收法和免疫亲和层析法,从艾氏腹水癌患鼠腹水DNA结合蛋白中,分离得到了一种高分子量DNA结合蛋白。在免疫双扩散反应中,它与抗艾氏腹水癌患鼠血清DNA结合蛋白的兎抗血清反应形成一条沉淀线,但与正常小鼠血清DNA结合蛋白的兎抗血清不形成沉淀线。该DNA结合蛋白样品用2-巯基乙醇还原后,经SDC-PAGE分析,测得其分子量约为41000。     

Circulating immune complexes and in vitro cell reactivity in paracoccidioidomycosis

The severest forms of paracoccidioidomycosis (Pcm) are associated with impaired cell-mediated immunity, a phenomenon that is reversible with therapy. It has been postulated that plasma factors could be responsible for such immune dysfunction. In this report, circulating immune complexes (CIC) were measured by the Raji cell radioimmunoassay (Raji) and by the¹²⁵I-C1q binding assay (C1q-BA) in sera from 14 patients with either active or inactive forms of Pcm and from 15 healthy controls. The C1q-B A revealed significantly elevated levels of CIC in the sera of all but one of the patients. Four of the 8 active (62%) and 2 of the 6 inactive (33%) patients had CIC levels significantly higher than the controls as determined by the Raji test. Significantly increased levels of CIC were detected only in the active patients by the Raji test. The serum of one of the patients, with a generalized infection and depressed lymphocyte responsiveness, was examined and found to contain a factor which depressed the in vitro proliferation of both homologous and normal lymphocytes. We also found that pre-culture of the patients' lymphocytes before stimulation restored their proliferative capacity, and IC were detectable in the culture supernatants. However, the subsequent addition of the patients' serum to such precultured cells did not reinduce the depression. It is suggested therefore, that the depression of T cell responses observed in Pcm is due to the presence of IC which may interact reversibly with the responding cells and/or activate a suppressor cell population whose activity is diminished by preculture.     

流行性出血热循环免疫复合物组分分析

应用ＳＤＳ－聚丙烯酰胺凝胶电泳及免疫转印技术对流行性出血热患才血清中免疫合物组分进行了分析。流行性出血热循环免疫复合物经ＳＤＳ－ＰＡＧＥ分离,考马斯亮兰染色,显色主要有７条带,分子量分别为２３ｋＤ,５０ｋＤ,５２ｋＤ,６５ｋＤ,７２ｋＤ,８０ｋＤ及１００ｋＤ。采用该病毒特异性抗血清、单克隆抗体以及人免疫球蛋白、补体成分抗血清识别,在其特环免疫复合物中可检出特异性病毒抗在及相应的免疫球状蛋白和补体成     

The development of a C1q-solid-phase immunoenzyme method for determining circulating immune complexes]

The method of quantitative enzyme immunoassay (EIA) for the determination of circulating immune complexes (CIC) was developed on the basis of solid-phase human C1q. The calibration curve was plotted with the use of aggregated human gamma-globulin (AHGG), the optimum range of concentration being 15-500 microg/ml. In the process of approbation on clinical material the method revealed an elevated level of CIC in the sera of patients in comparison with their level in the sera of healthy donors. Out of 40 studied serum samples from patients with Yersinia infection, in 3 serum samples the levels of CIC was 26, 65 and 94 microg of AHGG equivalents per ml. In 4 out of 46 studied serum samples obtained from patients with diagnosed Yersinia arthritis the level of CIC was 12, 27, 46 and 186 microg of AHGG per ml, and in serum samples from healthy donors this level was 8.6 microg/ml [corrected].     

Evaluation of an enzyme-linked immunosorbent Raji cell assay (ELISA) in the investigation of gastrointestinal cancer

The levels of circulating immune complexes (CICs) have been estimated in a group of patients with colorectal cancer and gastric cancer, in addition to which a normal range has been established in a group of patients with benign gastrointestinal disease. A newly developed enzyme-linked immunosorbent Raji cell assay has been used in this study. Overall only 30% of patients with gastrointestinal cancer showed elevation of CIC levels outside the normal range. Elevated levels correlated with tumour differentiation bud did not correlate with site of disease or with the presence of metastases. In an attempt to define the specificity of CIC estimation, soluble tumour extract was added to sera from tumour-bearing patients. Specific IC elevations were produced by addition of allogeneic tumour extract of colon cancer in patients with colorectal cancer; this phenomenon was not seen when the same extract was added to the sera of patients with gastric cancer.     

Evaluation of an enzyme-linked immunosorbent Raji cell assay (ELISA) in the investigation of gastrointestinal cancer

Summary The levels of circulating immune complexes (CICs) have been estimated in a group of patients with colorectal cancer and gastric cancer, in addition to which a normal range has been established in a group of patients with benign gastrointestinal disease. A newly developed enzyme-linked immunosorbent Raji cell assay has been used in this study. Overall only 30% of patients with gastrointestinal cancer showed elevation of CIC levels outside the normal range. Elevated levels correlated with tumour differentiation bud did not correlate with site of disease or with the presence of metastases. In an attempt to define the specificity of CIC estimation, soluble tumour extract was added to sera from tumour-bearing patients. Specific IC elevations were produced by addition of allogeneic tumour extract of colon cancer in patients with colorectal cancer; this phenomenon was not seen when the same extract was added to the sera of patients with gastric cancer.     

Analysis of circulating immune complexes (CIC) in tuberculosis: levels of specific antibody and antigens in CIC and relationship with serum antibody

Eighty sera from tuberculosis (TB) patients, 16 Indian and 10 American control sera were analyzed by ELISA for relative titres of antibody against mycobacterial antigens. Levels of specific antibody and mycobacterial Ag in circulating immune complexes (CIC) isolated from these sera were also studied. All these parameters were found to be elevated in TB sera as compared to control sera. Maximum increase was however noted in CIC specific antibody titres. A good correlation was observed between serum and CIC levels of specific antibody (r = 0.72) and between specific antigen (Ag) and antibody (Ab) levels within CIC (r = 0.64). In a few of the TB sera examined, CIC specific Ab contributed less than 1% to the Ab titres in sera. In order to examine the differences between different subgroups within TB patients, a statistical analysis of variance was performed. Sex of the patients had no effect on any parameter. Sputum-positive patients had significantly higher levels of CIC Ag and Ab than the sputum-negative patients, although no significant difference occurred in respect to serum Ab. All three parameters were significantly higher in patients on chemotherapy as compared to fresh untreated cases. The relevance of these observations to the development of a CIC-based immunodiagnostic assay for TB is discussed.     

Presence of antibodies specifically reacting with structural proteins of the mammary cancer virus among antibodies found in circulating immune complexes in patients with breast cancer

Circulating immune complexes were precipitated from breast cancer patients' sera using 2.5% polyethylene glycol. CIC isolated from 70 ml of sera from 15 patients were dissociated and immunoglobulin-containing fraction was prepared by chromatography on Sephadex G-200 column. The fraction contained IgG specific for MuMTV structural proteins, as revealed by ELISA. CIC preparations from 22 sera of breast cancer patients were digested with pepsin; Fab' fragment preparations were also analysed by ELISA, only one of them was MuMTV-specific. IgG and Fab' fragments isolated from CIC reacted specifically with MuMTV proteins, the reaction was not blocked by virus-free murine milk or other retroviruses (Ra-MuLV and MPMV).     

High-titered anti-HBs plasma donors--another form of post-hepatitic immunopathy syndrome

In the population of 55 high-titered anti-HBs donors only 23 tolerated plasmapheretic collections without intermittent elevations or ALT activity. In 4 persons a RIA-detected HBsAg circulated along with high-titered anti-HBs. In 73.8% of donors anti-HBs was accompanied by an anti-HBe antibody which also appeared in the HBIG preparation HEPAGA and can perhaps participate on its protective influence. Circulating immune complexes (CIC) were detected in 89.1%. No HBsAg, HBeAg, or albumin were detected in CIC isolated from anti-HBs sera in spite of their content in CIC isolated from HBsAg carriers. Thus, CIC carriers found in normal population with a prevalence of 1.0% can be divided into 0.6% of HHsAg-containing CIC and 0.4% of HBsAg-lacking CIC carriers with anti-HBs attesting the hepatitic origin in a considerable part of them. The continuing production of alienated CIC-forming antigens and a common origin combine these two forms of post-hepatitic development to a syndrome of post-hepatitic immunopathy which seems to be the most frequent source of CIC in a normal population. All the donors and HEPAGA were anti-HBc positive, as well, but this antibody possessed the IgM character only in 4.3% of the donors. Mean serum ferritin levels in the anti-HBs donors were distinctly higher than those found in normal populations of both men and women but the differences were statistically not significant due to high variability.